Use of Ion Exchange Membranes for Selective Recovery of Aspergillus awamori Glucoamylase and Phage T4 Lysozyme

Our research examined whether enhanced separation and purification of mutant enzymes could be obtained on ion-exchange membranes. Solutions of three mutants of Aspergillus awamori glucoamylase were passed through an anionic exchange membrane, as well as one mutant of T4 lysozyme through a cationic exchange membrane. The mutant enzymes were modified by adding charged fusions\u27\u27, polypeptides of either aspartic acid residues to increase the overall negative charge of the enzyme or arginine residues to increase the overall positive charge. The effect of the mutations on the purification of glucoamylase from a modified fermentation broth were examined at two different elution pHs, 4.5 and 6.0. The use of the charged fusions provided significantly improved purification capabilities over control versions. Both the small scale glucoamylase runs and the scaled up experiments had overall purification factors of around two, with a peak purification factor of near 7 for GA\u27CDlO. Elution of glucoamylase at pH 4.5 did not lead to an increase in separability as compared to that obtained at pH 6.0. Initial trials using a purified lysozyme mutant showed significant binding capabilities. Further experiments with this protein need to be done to determine scale up potential

Fatty acid nitroalkenes ameliorate glucose intolerance and pulmonary hypertension in high-fat diet-induced obesity

Aims Obesity is a risk factor for diabetes and cardiovascular diseases, with the incidence of these disorders becoming epidemic. Pathogenic responses to obesity have been ascribed to adipose tissue (AT) dysfunction that promotes bioactive mediator secretion from visceral AT and the initiation of pro-inflammatory events that induce oxidative stress and tissue dysfunction. Current understanding supports that suppressing pro-inflammatory and oxidative events promotes improved metabolic and cardiovascular function. In this regard, electrophilic nitro-fatty acids display pleiotropic anti-inflammatory signalling actions. Methods and results It was hypothesized that high-fat diet (HFD)-induced inflammatory and metabolic responses, manifested by loss of glucose tolerance and vascular dysfunction, would be attenuated by systemic administration of nitrooctadecenoic acid (OA-NO2). Male C57BL/6j mice subjected to a HFD for 20 weeks displayed increased adiposity, fasting glucose, and insulin levels, which led to glucose intolerance and pulmonary hypertension, characterized by increased right ventricular (RV) end-systolic pressure (RVESP) and pulmonary vascular resistance (PVR). This was associated with increased lung xanthine oxidoreductase (XO) activity, macrophage infiltration, and enhanced expression of pro-inflammatory cytokines. Left ventricular (LV) end-diastolic pressure remained unaltered, indicating that the HFD produces pulmonary vascular remodelling, rather than LV dysfunction and pulmonary venous hypertension. Administration of OA-NO2 for the final 6.5 weeks of HFD improved glucose tolerance and significantly attenuated HFD-induced RVESP, PVR, RV hypertrophy, lung XO activity, oxidative stress, and pro-inflammatory pulmonary cytokine levels. Conclusions These observations support that the pleiotropic signalling actions of electrophilic fatty acids represent a therapeutic strategy for limiting the complex pathogenic responses instigated by obesity.Fil: Kelley, Eric E.. University of Pittsburgh; Estados UnidosFil: Baust, Jeff. University of Pittsburgh; Estados UnidosFil: Bonacci, Gustavo Roberto. University of Pittsburgh; Estados Unidos. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; ArgentinaFil: Golin Bisello, Franca. University of Pittsburgh; Estados UnidosFil: Devlin, Jason E.. University of Pittsburgh; Estados UnidosFil: Croix, Claudette M. St.. University of Pittsburgh; Estados UnidosFil: Watkins, Simon C.. University of Pittsburgh; Estados UnidosFil: Gor, Sonia. University of Pittsburgh; Estados UnidosFil: Cantu Medellin, Nadiezhda. University of Pittsburgh; Estados UnidosFil: Weidert, Eric R.. University of Pittsburgh; Estados UnidosFil: Frisbee,Jefferson C.. University of Virginia; Estados UnidosFil: Gladwin, Mark T.. University of Pittsburgh; Estados UnidosFil: Champion, Hunter C.. University of Pittsburgh; Estados UnidosFil: Freeman, Bruce A.. University of Pittsburgh; Estados UnidosFil: Khoo, Nicholas K.H.. University of Pittsburgh; Estados Unido

REC, Drosophila MCM8, Drives Formation of Meiotic Crossovers

Crossovers ensure the accurate segregation of homologous chromosomes from one another during meiosis. Here, we describe the identity and function of the Drosophila melanogaster gene recombination defective (rec), which is required for most meiotic crossing over. We show that rec encodes a member of the mini-chromosome maintenance (MCM) protein family. Six MCM proteins (MCM2–7) are essential for DNA replication and are found in all eukaryotes. REC is the Drosophila ortholog of the recently identified seventh member of this family, MCM8. Our phylogenetic analysis reveals the existence of yet another family member, MCM9, and shows that MCM8 and MCM9 arose early in eukaryotic evolution, though one or both have been lost in multiple eukaryotic lineages. Drosophila has lost MCM9 but retained MCM8, represented by REC. We used genetic and molecular methods to study the function of REC in meiotic recombination. Epistasis experiments suggest that REC acts after the Rad51 ortholog SPN-A but before the endonuclease MEI-9. Although crossovers are reduced by 95% in rec mutants, the frequency of noncrossover gene conversion is significantly increased. Interestingly, gene conversion tracts in rec mutants are about half the length of tracts in wild-type flies. To account for these phenotypes, we propose that REC facilitates repair synthesis during meiotic recombination. In the absence of REC, synthesis does not proceed far enough to allow formation of an intermediate that can give rise to crossovers, and recombination proceeds via synthesis-dependent strand annealing to generate only noncrossover products

Stellar photometry with the Hubble Space Telescope Wide-field/Planetary camera - A progress report

We describe the prospects for the use of the Wide-Field/Planetary Camera (WFPC) for stellar photometry. The large halos of the point-spread function (PSF) resulting from spherical aberration and from spatial, temporal, and color variations of the PSF are the main limitations to accurate photometry. Degradations caused by crowding are exacerbated by the halos of the PSF. Here we attempt to quantify these effects and determine the current accuracy of stellar photometry with the WFPC. In realistic cases, the brighter stars in crowded fields have 0.09 mag errors; fainter stars have larger errors depending on the degree of crowding. We find that measuring Cepheids in Virgo Cluster galaxies is not currently possible without inordinate increases in exposure times

Reduction of PG:1115+080 Images

The data are three exposures in PC6 through F785LP obtained on March 3, 1991. The exposure times are 120, 400, and 400 seconds. The data are reduced with the "standard" WFPC reduction scheme: A-to-D correction, DC bias subtraction, AC bias subtraction, dark current subtraction, preflash subtraction, and flat field normalization, using the best available calibration data. The exposures are combined into a weighted average normalized to 400 seconds exposure time, so one DN (data number) is about 17.25 electrons. At this step, cosmic rays are removed by intercomparison of the three images

A Technique for Measuring Petal Gloss, with Examples from the Namaqualand Flora

The degree of floral gloss varies between species. However, little is known about this distinctive floral trait, even though it could be a key feature of floral biotic and abiotic interactions. One reason for the absence of knowledge is the lack of a simple, repeatable method of gloss measurement that can be used in the field to study floral gloss. A protocol is described for measuring gloss in petal samples collected in the field, using a glossmeter. Repeatability of the technique is assessed. We demonstrate a simple yet highly accurate and repeatable method that can easily be implemented in the field. We also highlight the huge variety of glossiness found within flowers and between species in a sample of spring-blooming flowers collected in Namaqualand, South Africa. We discuss the potential uses of this method and its applications for furthering studies in plant-pollinator interactions. We also discuss the potential functions of gloss in flowers

Masses, radii, and orbits of small Kepler planets : The transition from gaseous to rocky planets

We report on the masses, sizes, and orbits of the planets orbiting 22 Kepler stars. There are 49 planet candidates around these stars, including 42 detected through transits and 7 revealed by precise Doppler measurements of the host stars. Based on an analysis of the Kepler brightness measurements, along with high-resolution imaging and spectroscopy, Doppler spectroscopy, and (for 11 stars) asteroseismology, we establish low false-positive probabilities (FPPs) for all of the transiting planets (41 of 42 have an FPP under 1%), and we constrain their sizes and masses. Most of the transiting planets are smaller than three times the size of Earth. For 16 planets, the Doppler signal was securely detected, providing a direct measurement of the planet's mass. For the other 26 planets we provide either marginal mass measurements or upper limits to their masses and densities; in many cases we can rule out a rocky composition. We identify six planets with densities above 5 g cm-3, suggesting a mostly rocky interior for them. Indeed, the only planets that are compatible with a purely rocky composition are smaller than 2 R ⊕. Larger planets evidently contain a larger fraction of low-density material (H, He, and H2O).Peer reviewedFinal Accepted Versio

Kate 2012

Each year, kate seeks to: explore ideas about normative gender, sex, and sexuality work against oppression and hierarchies of power in any and all forms serve as a voice for race and gender equity as well as queer positivity encourage the silent to speak and feel less afraid build a zine and community that we care about and trusthttps://digitalcommons.otterbein.edu/kate/1007/thumbnail.jp